SOLIScript 1-step Multiplex Probe Kit is optimized for probe based one-step RT real-time quantitative PCR assays. It contains all the components necessary (except template, primers, and probes) to perform cDNA synthesis and probe based qPCR in a single tube. SOLIScript RT is an in silico-engineered thermostable reverse transcriptase that is active at higher temperatures (up to 60C) for providing highly specific results and working with templates with high secondary structure. HOT FIREPol® DNA Polymerase is activated by a 10 min incubation step at 95°C. This prevents extension of nonspecifically annealed primers and primer-dimers formed at low temperatures during qPCR setup. SOLIScript 1-step Multiplex Probe Kit is optimized for DNA/LNA hydrolysis probes based on the 5' flap endonuclease activity.
• Detection and quantification of RNA targets with qPCR
• DNA/LNA hydrolysis probe based assays
• Suitable for singleplex and multiplex reactions
• 5x One-step Multiplex Probe Mix HOT FIREPol® DNA polymerase, reaction buffer, dNTPs (including dUTP), 15 mM MgCl2 (1x qPCR solution - 3 mM MgCl2)
• 40x One-step SOLIScript Mix SOLIScript Reverse Transcriptase, RiboGrip RNase Inhibitor
• Water, nuclease free
Shipping and Storage conditions: Routine storage: -20C Shipping and temporary storage for up to 1 month at room temperature has no detrimental effects on the quality of SOLIScript 1-step Multiplex Probe Kit.
Recommended RT-qPCR reaction mix:
Component 20 µl/rxn Final conc.
5x One-step Multiplex Probe Mix 4 µl
1x 40x One-step SOLIScript Mix 0.5 µl
1x Primer Forward (10 pmol/µl) 0.4-0.8 µl 200-400 nM Primer Reverse (10 pmol/µl) 0.4-0.8 µl 200-400 nM Probe Variable 100-250 nM
Optional: thermolabile UDG Variable Variable RNA template (1 pg - 1µg )
Variable Water, nuclease free up to 20 µl Total 20 µl
Recommended RT-qPCR cycles:
Cycle step Temp. Time Cycles Reverse transcription 50-60C 15 min 1 Initial activation 95C 10 min 1 Denaturation 95C 15 s 40 Annealing/Elongation 60C 60 s
Safety warnings and precautions: This product and its components should be handled only by persons trained in laboratory techniques. It is advisable to wear suitable protective clothing, such as laboratory overalls, gloves and safety glasses. Care should be taken to avoid contact with skin or eyes. In case of contact with skin or eyes, wash imme